酸性铁蛋白放免分析中抗体纯化的可行性  

STUDY OF THE FEASIBILITY OF ANTIBODY PURIFICATION IN ACIDIC ISOFERRITIN RADIOIOIMMUNOASSSAY

作　　者：李文新[1] 王桂莲[1] 黄飙[1] 张蓬芬 陶永辉[1] 金坚[1] 

机构地区：[1]江苏省原子医学研究所

出　　处：《同位素》1996年第3期145-150,共6页Journal of Isotopes

摘　　要：用改进的工艺自人心肌组织中提取的高纯度酸性铁蛋白（ＡＩＦ），与经溴化氰（ＢｒＣＮ）活化的ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ偶联，制得ＡＩＦ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ亲和层析柱。用经该柱纯化的兔抗ＡＩＦ抗血清来组建酸性铁蛋白放免分析法（ＡＩＦＲＩＡ）用四参数Ｌｏｇｉｓｔｉｃ数据处理程序处理数据。批内、批间ＣＶ分别为１．６５％和９．７１％，灵敏度为１．８９μｇ／ｌ，回收率为１０２％，可测范围为７．０—３６９．６μｇ／ｌ，ＥＤ５０为２７．５０μｇ／ｌ，与ＡＦＰ、ＣＥＡ、ＬＦ无交叉反应，同常规铁蛋白的交叉反应率为１．７４％。对照组男、女性各３０例，血清ＡＩＦ含量（ｘ±２ｓ）分别为１６．７６±６．９８μｇ／ｌ和１３．３２±６．２５μｇ／１，差异不显著（Ｐ＜０．０５）。１０例肝癌患者血清ＡＩＦ含量为３６．０９±１５．８４μｇ／ｌ，较对照组有非常显著的增高（Ｐ＜０．００１）。上述结果提示，用亲和层析技术纯化ＡＩＦ抗血清是可行的，有利于提高ＲＩＡ的灵敏度、特异性、准确性及临床有效性。The high purity acidic isoferritin (AIF) extracted from the human heart tissue with a medifiedtechnology is coupled to cyanogen bromide-activated Sepharoce CL-6B to preP8re AIF-Sepharose CL6B affinity chromatiographic column. The rabbit anti-AIF serum purified from this column is used toestablish a new AIF RIA. The data obtained are processed by the 4-PL method. T'he intra- and interCV are 1. 65% and 9. 71 %, respectively. The sensitivity, ED50, recovery rate and effective assayrange are 1. 89μg/l, 27. 50 μg/l, 102% and 7. 0-369. 6μg/l, respetively. No cross-reaction withAFP、CEA and LF is oberved and the cross-reaction with ferritin is only 1. 74 %. The serum AIF levels in 30 male or female normal controls are 16. 76±6. 98μg/l and 13. 32±6. 25μg/l, respectively. No statistical difference is observed(P>0. 05). But the serum AIF levels in 10 patients ofhepotocarcinoma are 36. 09±15. 84 μg/l. being strikingly higher than those of normal controls (P<0. 001 ). This suggests that the method of purifying the rabbit anti-AIF serum by affinity chromatography is fasiible and will help to improve the sensitivity, spectificity, accuracy and clinical effectivenessof the AIF RIA.

关 键 词：酸性铁蛋白 放射免疫分析 亲和层析 交叉反应 肝癌 

分 类 号：R817.4[医药卫生—影像医学与核医学] R446.6[医药卫生—放射医学]